Genetic characterisation of Staphylococcus aureus isolated from milk and nasal samples of healthy cows in Tunisia: First report of ST97-t267-agrI-SCCmecV MRSA of bovine origin in Tunisia.

This study aimed to screen for and characterise methicillin-susceptible Staphylococcus aureus (MSSA) and methicillin-resistant S. aureus (MRSA) in nasal swabs and milk from healthy cows from different regions in Tunisia. A total of 141 Staphylococcus spp. isolates were recovered from milk and nasal samples of cows. S. aureus isolates were further characterised by determining their antimicrobial susceptibilities, genes encoding antimicrobial resistance and virulence factors, biofilm production, agr type and PFGE. spa and SCCmec typing and MLST were also performed for the MRSA isolate. Twenty-seven isolates (19.1%) were identified as S. aureus, of which 26 were MSSA and 1 was MRSA. The MSSA isolates were resistant to penicillin (73.1%), fusidic acid (61.5%), clindamycin (34.6%) and erythromycin (34.6%). The MRSA isolate, from a milk sample, was resistant to cefoxitin, penicillin, fusidic acid, amikacin and clindamycin. Twenty-five isolates (92.6%) had at least one enterotoxin gene. Only four isolates (14.8%) were positive for the tsst-1 gene. Genes encoding the exfoliative toxins D and A were detected in 9 (33.3%) and 6 (22.2%) isolates, respectively. The single MRSA isolate and 22 MSSA isolates were biofilm-producers on Congo red agar plates. Twelve pulsotypes were identified amongst 25 MSSA isolates revealing the clonal diversity of these isolates; however, one MSSA isolate was identified as CC398. The MRSA isolate was PVL-negative and was typed as ST97-t267-agrI-SCCmecV. Contamination of milk with S. aureus, especially enterotoxin- and TSST-1-positive strains, poses a potential public-health threat. This is the first report of MRSA of bovine origin in Tunisia.

Khemiri M ，Abbassi MS ，Couto N ，Mansouri R ，Hammami S ，Pomba C ... -  《-》

Molecular Characterization and Clonal Diversity of Methicillin-Resistant and -Susceptible Staphylococcus aureus Isolates of Milk of Cows with Clinical Mastitis in Tunisia.

Klibi A ，Jouini A ，Gómez P ，Slimene K ，Ceballos S ，Torres C ，Maaroufi A ... -  《-》

Staphylococcus aureus nasal carriage, virulence traits, antibiotic resistance mechanisms, and genetic lineages in healthy humans in Spain, with detection of CC398 and CC97 strains.

S. aureus nasal carriage was investigated in 278 healthy humans, determining the antibiotic resistance mechanisms, virulence traits, and genetic lineages of recovered isolates. Nasal samples were cultured in specific media for S. aureus and methicillin-resistant S. aureus (MRSA) recovery. S. aureus was detected in 53 of 278 nasal samples (19.1%): MRSA was found in one sample (0.4%) and methicillin-susceptible S. aureus (MSSA) in the remaining 52 samples. The MRSA isolate was typed as ST1649-t701-agrI-SCCmec-IVc and only exhibited resistance to beta-lactams. A high diversity of spa types (n=37) was identified among the 52 MSSA, identifying 5 new spa-types. Multi-locus sequence typing (MLST) typing was performed in 30 selected MSSA, detecting 16 different sequence types, 2 of them being new. MSSA strains presented agr types I (30.2%), II (30.2%), III (34%), and IV (5.6%). Eleven strains showed erythromycin resistance and harbored different combinations of erm(A), erm(B), erm(C), erm(T), and msr(A) genes. Two strains exhibited ciprofloxacin resistance, and one of them presented amino acid changes in GyrA and GrlA proteins. The presence of 28 genes encoding staphylococcal toxins was investigated by PCR in all 53 S. aureus isolates. The toxic shock syndrome toxin 1 (TSST-1) gene was detected in 15 MSSA isolates (11 of them typed within the clonal complex CC30) and the gene of exfoliative toxin A in 2 strains. Different combinations of enterotoxin genes were identified among S. aureus strains. None of the S. aureus isolates harbored the Panton-Valentine leukocidin gene. Two MSSA presented the sequence-type ST398 [harboring erm(T) gene], and 2 additional isolates were typed as ST97. Interestingly, MSSA CC398 and CC97 isolates were detected. These clonal complexes are associated with food-producing animals.

Lozano C ，Gómez-Sanz E ，Benito D ，Aspiroz C ，Zarazaga M ，Torres C ... -  《-》

Prevalence, antibiotic resistance, virulence traits and genetic lineages of Staphylococcus aureus in healthy sheep in Tunisia.

Nasal swabs of 163 healthy sheep were obtained from two farms and one abattoir in Tunisia during 2010. Samples were inoculated in Baird Parker agar and ORSAB medium for Staphylococcus aureus and methicillin-resistant S. aureus (MRSA) recovery, respectively. MRSA was detected in 5 of these 163 samples (3%) in ORSAB medium, and one isolate per sample was further studied. MRSA isolates were mecA-positive, typed as ST153-CC80-t044-agrIII, and contained blaZ, ant(6)-Ia, aph(3')-IIIa, erm(C), tet(K), and fusB genes encoding penicillin, streptomycin, kanamycin, erythromycin, tetracycline and fusidic acid resistance, respectively. These MRSA isolates showed indistinguishable or closely related PFGE-patterns and harboured the lukF/lukS gene encoding the Panton-Valentine leukocidin and the luk-ED, hla, hld, and hlg(v) genes. Methicillin-susceptible S. aureus (MSSA) were recovered in 68 of the 163 samples (41.7%) and one isolate per sample was characterized. Most of MSSA (82.4%) showed susceptibility to the tested antibiotics with exceptions: penicillin (6%, carrying blaZ gene), tetracycline (19%, carrying tet(K) gene) and fusidic acid (9%). The following toxin-genes were identified among MSSA: tst (53 isolates), luk-M (52), luk-ED, hla, hlb, hld and hlg(v) (67), hlg (1), sec (49), sel (52), and the egc-cluster-like sen-sem-sei-seo-seg (1). Ten spa-types (two of them new ones) and nine sequence types (six new ones) were detected among the 73 S. aureus isolates, and they were ascribed to agr types I and III. All MRSA and MSSA isolates were able to coagulate bovine plasma and MRSA harboured the immune-evasion-gene-cluster type E. Conclusions. Nares of healthy sheep could be a reservoir of PVL-positive community-associated-MRSA and also of TSST-positive S. aureus isolates, with potential implications in public health.

Gharsa H ，Ben Slama K ，Lozano C ，Gómez-Sanz E ，Klibi N ，Ben Sallem R ，Gómez P ，Zarazaga M ，Boudabous A ，Torres C ... -  《-》

Antimicrobial resistance, virulence gene profile and molecular typing of Staphylococcus aureus isolates from dairy cows in Xinjiang Province, northwest China.

Staphylococcus aureus (SA) is a major pathogen causing dairy cow mastitis and endometritis. Recently, animal-derived SA strains pose a serious public-health threat. However, little is known about antimicrobial resistance and virulence factors of SA isolated from dairy cows in Xinjiang, China. In this study, antimicrobial resistance, virulence gene profiles and genotypes of SA from clinical mastitis and endometritis in dairy cows were investigated. A total of 337 clinical samples (186 milk samples from clinical mastitis cases and 151 endometritis swab samples) were collected from 15 large-scale dairy farms and were screened for SA. All SA isolates were subjected to antimicrobial susceptibility testing, detection of virulence genes and molecular typing. A total of 155 SA strains were isolated; 22 (14.2%) were methicillin-resistant S. aureus (MRSA). Resistance of MRSA isolates was significantly higher than that of methicillin-susceptible S. aureus (MSSA). The percentage of virulence genes varied between MSSA and MRSA. The strains could be divided into two SCCmec types (I and IVa), three agr types (I, II and III) and four spa types (t779, t2883, t13751 and t1939). MLST identified 14 sequence types, among which ST1 and ST9 had relatively high detection rates. These findings revealed that ST9-t1939-agrI was the main genotype of MSSA, whilst ST1-SCCmecI-t1939-agrI was the main genotype of MRSA from dairy cows. More significantly, a novel ST (STX) was identified for the first time. The majority of SA strains from dairy cows were multidrug-resistant and carried multiple virulence genes, posing a potential public-health risk.

Dan M ，Yehui W ，Qingling M ，Jun Q ，Xingxing Z ，Shuai M ，Kuojun C ，Jinsheng Z ，Zibing C ，Zaichao Z ，Xuepeng C ... -  《-》